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Characterizing yeast promoters used in Kluyveromyces marxianus.
World J Microbiol Biotechnol. 2015 Oct; 31(10):1641-6.WJ

Abstract

Fermentation at higher temperatures can potentially reduce the cooling cost in large-scale fermentation and reduce the contamination risk. Thus, the thermotolerant yeast, Kluyveromyces marxianus, which can grow and ferment at elevated temperatures, is a promising biotechnological tool for future applications. However, the promoters used in K. marxianus are not well characterized, especially at elevated temperatures, which is important in efficient metabolic pathway construction. In this study, six constitutive promoters (P(TDH3), P(PGK), and P(ADH1) from both Saccharomyces cerevisiae and K. marxianus) were evaluated in K. marxianus through the heterologous expression of the KlLAC4, GUSA, and SH BLE genes at various temperatures, with various carbon sources and oxygen conditions. The expression was evaluated at the transcription and protein level using real-time PCR and protein activity determination to eliminate the effect of heterologous protein stability. While the transcription of all the promoters decreased at higher temperatures, the order of their promoting strength at various temperatures with glucose as the carbon source was P(KmPGK) > P(KmTDH3) > P(ScPGK) > P(ScTDH3) > P(KmADH1) > P(ScADH1). When glycerol or xylose was supplied as the carbon source at 42 °C, the order of promoter strength was P(KmPGK) > P(ScPGK) > P(KmADH1) > P(ScADH1) > P(ScTDH3) > P(KmTDH3). The promoter activity of P TDH3 decreased significantly, while the promoter activity of both of the P(ADH1) promoters increased. Oxygen conditions had non-significant effect. The results of this study provide important information for fine-tuned pathway construction for the metabolic engineering of K. marxianus.

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Authors+Show Affiliations

Yang C
School of Life Science, University of Science and Technology of China, Hefei, 230027, Anhui, People's Republic of China. Hefei National Laboratory for Physical Science at the Microscale, University of Science and Technology of China, Hefei, 230026, Anhui, People's Republic of China.
Hu S
School of Life Science, University of Science and Technology of China, Hefei, 230027, Anhui, People's Republic of China. Hefei National Laboratory for Physical Science at the Microscale, University of Science and Technology of China, Hefei, 230026, Anhui, People's Republic of China.
Zhu S
School of Life Science, University of Science and Technology of China, Hefei, 230027, Anhui, People's Republic of China. Hefei National Laboratory for Physical Science at the Microscale, University of Science and Technology of China, Hefei, 230026, Anhui, People's Republic of China.
Wang D
School of Life Science, University of Science and Technology of China, Hefei, 230027, Anhui, People's Republic of China. Hefei National Laboratory for Physical Science at the Microscale, University of Science and Technology of China, Hefei, 230026, Anhui, People's Republic of China.
Gao X
School of Life Science, University of Science and Technology of China, Hefei, 230027, Anhui, People's Republic of China. Hefei National Laboratory for Physical Science at the Microscale, University of Science and Technology of China, Hefei, 230026, Anhui, People's Republic of China. Department of Biology and Biochemistry, University of Houston, Houston, 77004-5001, USA.
Hong J
School of Life Science, University of Science and Technology of China, Hefei, 230027, Anhui, People's Republic of China. hjiong@ustc.edu.cn. Hefei National Laboratory for Physical Science at the Microscale, University of Science and Technology of China, Hefei, 230026, Anhui, People's Republic of China. hjiong@ustc.edu.cn.

MeSH

Gene ExpressionGene Expression ProfilingKluyveromycesPromoter Regions, GeneticRecombinant ProteinsSaccharomyces cerevisiaeTemperature

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

26164057

Citation

Yang, Chun, et al. "Characterizing Yeast Promoters Used in Kluyveromyces Marxianus." World Journal of Microbiology & Biotechnology, vol. 31, no. 10, 2015, pp. 1641-6.
Yang C, Hu S, Zhu S, et al. Characterizing yeast promoters used in Kluyveromyces marxianus. World J Microbiol Biotechnol. 2015;31(10):1641-6.
Yang, C., Hu, S., Zhu, S., Wang, D., Gao, X., & Hong, J. (2015). Characterizing yeast promoters used in Kluyveromyces marxianus. World Journal of Microbiology & Biotechnology, 31(10), 1641-6. https://doi.org/10.1007/s11274-015-1899-x
Yang C, et al. Characterizing Yeast Promoters Used in Kluyveromyces Marxianus. World J Microbiol Biotechnol. 2015;31(10):1641-6. PubMed PMID: 26164057.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Characterizing yeast promoters used in Kluyveromyces marxianus. AU - Yang,Chun, AU - Hu,Shenglin, AU - Zhu,Songli, AU - Wang,Dongmei, AU - Gao,Xiaolian, AU - Hong,Jiong, Y1 - 2015/07/12/ PY - 2015/04/12/received PY - 2015/07/06/accepted PY - 2015/7/13/entrez PY - 2015/7/15/pubmed PY - 2016/6/2/medline KW - Carbon source KW - Elevated temperature KW - Kluyveromyces marxianus KW - Promoter SP - 1641 EP - 6 JF - World journal of microbiology & biotechnology JO - World J Microbiol Biotechnol VL - 31 IS - 10 N2 - Fermentation at higher temperatures can potentially reduce the cooling cost in large-scale fermentation and reduce the contamination risk. Thus, the thermotolerant yeast, Kluyveromyces marxianus, which can grow and ferment at elevated temperatures, is a promising biotechnological tool for future applications. However, the promoters used in K. marxianus are not well characterized, especially at elevated temperatures, which is important in efficient metabolic pathway construction. In this study, six constitutive promoters (P(TDH3), P(PGK), and P(ADH1) from both Saccharomyces cerevisiae and K. marxianus) were evaluated in K. marxianus through the heterologous expression of the KlLAC4, GUSA, and SH BLE genes at various temperatures, with various carbon sources and oxygen conditions. The expression was evaluated at the transcription and protein level using real-time PCR and protein activity determination to eliminate the effect of heterologous protein stability. While the transcription of all the promoters decreased at higher temperatures, the order of their promoting strength at various temperatures with glucose as the carbon source was P(KmPGK) > P(KmTDH3) > P(ScPGK) > P(ScTDH3) > P(KmADH1) > P(ScADH1). When glycerol or xylose was supplied as the carbon source at 42 °C, the order of promoter strength was P(KmPGK) > P(ScPGK) > P(KmADH1) > P(ScADH1) > P(ScTDH3) > P(KmTDH3). The promoter activity of P TDH3 decreased significantly, while the promoter activity of both of the P(ADH1) promoters increased. Oxygen conditions had non-significant effect. The results of this study provide important information for fine-tuned pathway construction for the metabolic engineering of K. marxianus. SN - 1573-0972 UR - https://www.unboundmedicine.com/medline/citation/26164057/Characterizing_yeast_promoters_used_in_Kluyveromyces_marxianus_ DB - PRIME DP - Unbound Medicine ER -